Limits...
Urotensin II Protects Cardiomyocytes from Apoptosis Induced by Oxidative Stress through the CSE/H2S Pathway.

Gong H, Chen Z, Zhang X, Li Y, Zhang J, Chen Y, Ding Y, Zhang G, Yang C, Zhu Y, Zou Y - Int J Mol Sci (2015)

Bottom Line: UII pretreatment significantly reduced the number of apoptotic cardiomyocytes induced by H2O2; and it partly abolished the increase of pro-apoptotic protein Bax and the decrease of anti-apoptotic protein Bcl-2 in cardiomyocytes induced by H2O2.SiRNA targeted to the urotensin II receptor (UT) greatly inhibited these effects.In conclusion, UII rapidly promoted the phosphorylation of ERK and upregulated CSE level and H2S production, which in turn activated ERK signaling to protect cardiomyocytes from apoptosis under oxidative stress.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China. ghui1975@163.com.

ABSTRACT
Plasma urotensin II (UII) has been observed to be raised in patients with acute myocardial infarction; suggesting a possible cardiac protective role for this peptide. However, the molecular mechanism is unclear. Here, we treated cultured cardiomyocytes with H2O2 to induce oxidative stress; observed the effect of UII on H2O2-induced apoptosis and explored potential mechanisms. UII pretreatment significantly reduced the number of apoptotic cardiomyocytes induced by H2O2; and it partly abolished the increase of pro-apoptotic protein Bax and the decrease of anti-apoptotic protein Bcl-2 in cardiomyocytes induced by H2O2. SiRNA targeted to the urotensin II receptor (UT) greatly inhibited these effects. Further analysis revealed that UII increased the production of hydrogen sulfide (H2S) and the level of cystathionine-γ-lyase (CSE) by activating the ERK signaling in H2O2-treated-cardiomyocytes. Si-CSE or ERK inhibitor not only greatly inhibited the increase in CSE level or the phosphorylation of ERK induced by UII but also reversed anti-apoptosis of UII in H2O2-treated-cadiomyocytes. In conclusion, UII rapidly promoted the phosphorylation of ERK and upregulated CSE level and H2S production, which in turn activated ERK signaling to protect cardiomyocytes from apoptosis under oxidative stress. These results suggest that increased plasma UII level may protect cardiomyocytes at the early-phase of acute myocardial infarction in patients.

No MeSH data available.


Related in: MedlinePlus

UII induces the reciprocal regulation of CSE and ERK in cardiomyocytes exposed to H2O2. Representative image of p-ERK, ERK, CSE (A) and quantitative analysis of p-ERK/ERK level (B) by Western blot analysis in cultured cardiomyocytes. Cardiomyocytes were transfected with si-scramble or si-CSE for 48 h, treated with UII (0.1 μM) or PBS for 30 min, then exposed to 100 μM H2O2 for 10 min, analyzed for p-ERK and ERK levels, or exposed to 100 μM H2O2 for 24 h, and analyzed for CSE level. Values are mean ± SEM. **p < 0.01, UII + si-scram vs. Si-scram or UII + si-CSE vs. Si-CSE. #p < 0.05, UII + si-CSE vs. UII + si-scram. Representative image of p-ERK, ERK, CSE (C) and quantitative analysis of p-ERK/ERK level (D) by Western blot analysis in cultured cardiomyocytes. Cultured cardiomyocytes were treated with DMSO or U0126 with or without UII for 30 min, then exposed to 100 μM H2O2 for 10 min, analyzed for p-ERK and ERK level, or exposed to 100 μM H2O2 for 24 h, and analyzed for CSE level. Values are mean ± SEM. *p < 0.05, UII + DMSO vs. DMSO or UII + U0126 vs. UII + DMSO. #p < 0.05, UII + U0126 vs. UII + DMSO. All experiments were repeated independently at least three times.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4490456&req=5

ijms-16-12482-f004: UII induces the reciprocal regulation of CSE and ERK in cardiomyocytes exposed to H2O2. Representative image of p-ERK, ERK, CSE (A) and quantitative analysis of p-ERK/ERK level (B) by Western blot analysis in cultured cardiomyocytes. Cardiomyocytes were transfected with si-scramble or si-CSE for 48 h, treated with UII (0.1 μM) or PBS for 30 min, then exposed to 100 μM H2O2 for 10 min, analyzed for p-ERK and ERK levels, or exposed to 100 μM H2O2 for 24 h, and analyzed for CSE level. Values are mean ± SEM. **p < 0.01, UII + si-scram vs. Si-scram or UII + si-CSE vs. Si-CSE. #p < 0.05, UII + si-CSE vs. UII + si-scram. Representative image of p-ERK, ERK, CSE (C) and quantitative analysis of p-ERK/ERK level (D) by Western blot analysis in cultured cardiomyocytes. Cultured cardiomyocytes were treated with DMSO or U0126 with or without UII for 30 min, then exposed to 100 μM H2O2 for 10 min, analyzed for p-ERK and ERK level, or exposed to 100 μM H2O2 for 24 h, and analyzed for CSE level. Values are mean ± SEM. *p < 0.05, UII + DMSO vs. DMSO or UII + U0126 vs. UII + DMSO. #p < 0.05, UII + U0126 vs. UII + DMSO. All experiments were repeated independently at least three times.

Mentions: In order to detect the relationship between the increased CSE expression and ERK phosphorylation induced by UII, cardiomyocytes were transfected with siRNA targeted CSE (si-CSE), or treated with the ERK inhibitor, 1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene (U0126). P-ERK level and CSE levels were detected. Si-CSE effectively mediated knockdown of the expression of CSE in cultured cardiomyocytes, and it greatly inhibited the activation of ERK induced by UII in cardiomyocytes exposed to H2O2 (Figure 4A,C). ERK, one of the MAPK superfamily, is involved in cellular survival and apoptotic process [45]. Here, we provide evidence that UII-induced-activation of ERK was inhibited by si-CSE in H2O2-treated-cardiomyocytes. It is reasoned that ERK activation is involved in CSE/H2S-mediated-anti-apoptosis in cardiomyocytes under oxidative stress. These findings are in line with a previous study that CSE overexpression mediated protective effects associated with an increased ERK activation in VSMCs [46].


Urotensin II Protects Cardiomyocytes from Apoptosis Induced by Oxidative Stress through the CSE/H2S Pathway.

Gong H, Chen Z, Zhang X, Li Y, Zhang J, Chen Y, Ding Y, Zhang G, Yang C, Zhu Y, Zou Y - Int J Mol Sci (2015)

UII induces the reciprocal regulation of CSE and ERK in cardiomyocytes exposed to H2O2. Representative image of p-ERK, ERK, CSE (A) and quantitative analysis of p-ERK/ERK level (B) by Western blot analysis in cultured cardiomyocytes. Cardiomyocytes were transfected with si-scramble or si-CSE for 48 h, treated with UII (0.1 μM) or PBS for 30 min, then exposed to 100 μM H2O2 for 10 min, analyzed for p-ERK and ERK levels, or exposed to 100 μM H2O2 for 24 h, and analyzed for CSE level. Values are mean ± SEM. **p < 0.01, UII + si-scram vs. Si-scram or UII + si-CSE vs. Si-CSE. #p < 0.05, UII + si-CSE vs. UII + si-scram. Representative image of p-ERK, ERK, CSE (C) and quantitative analysis of p-ERK/ERK level (D) by Western blot analysis in cultured cardiomyocytes. Cultured cardiomyocytes were treated with DMSO or U0126 with or without UII for 30 min, then exposed to 100 μM H2O2 for 10 min, analyzed for p-ERK and ERK level, or exposed to 100 μM H2O2 for 24 h, and analyzed for CSE level. Values are mean ± SEM. *p < 0.05, UII + DMSO vs. DMSO or UII + U0126 vs. UII + DMSO. #p < 0.05, UII + U0126 vs. UII + DMSO. All experiments were repeated independently at least three times.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490456&req=5

ijms-16-12482-f004: UII induces the reciprocal regulation of CSE and ERK in cardiomyocytes exposed to H2O2. Representative image of p-ERK, ERK, CSE (A) and quantitative analysis of p-ERK/ERK level (B) by Western blot analysis in cultured cardiomyocytes. Cardiomyocytes were transfected with si-scramble or si-CSE for 48 h, treated with UII (0.1 μM) or PBS for 30 min, then exposed to 100 μM H2O2 for 10 min, analyzed for p-ERK and ERK levels, or exposed to 100 μM H2O2 for 24 h, and analyzed for CSE level. Values are mean ± SEM. **p < 0.01, UII + si-scram vs. Si-scram or UII + si-CSE vs. Si-CSE. #p < 0.05, UII + si-CSE vs. UII + si-scram. Representative image of p-ERK, ERK, CSE (C) and quantitative analysis of p-ERK/ERK level (D) by Western blot analysis in cultured cardiomyocytes. Cultured cardiomyocytes were treated with DMSO or U0126 with or without UII for 30 min, then exposed to 100 μM H2O2 for 10 min, analyzed for p-ERK and ERK level, or exposed to 100 μM H2O2 for 24 h, and analyzed for CSE level. Values are mean ± SEM. *p < 0.05, UII + DMSO vs. DMSO or UII + U0126 vs. UII + DMSO. #p < 0.05, UII + U0126 vs. UII + DMSO. All experiments were repeated independently at least three times.
Mentions: In order to detect the relationship between the increased CSE expression and ERK phosphorylation induced by UII, cardiomyocytes were transfected with siRNA targeted CSE (si-CSE), or treated with the ERK inhibitor, 1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene (U0126). P-ERK level and CSE levels were detected. Si-CSE effectively mediated knockdown of the expression of CSE in cultured cardiomyocytes, and it greatly inhibited the activation of ERK induced by UII in cardiomyocytes exposed to H2O2 (Figure 4A,C). ERK, one of the MAPK superfamily, is involved in cellular survival and apoptotic process [45]. Here, we provide evidence that UII-induced-activation of ERK was inhibited by si-CSE in H2O2-treated-cardiomyocytes. It is reasoned that ERK activation is involved in CSE/H2S-mediated-anti-apoptosis in cardiomyocytes under oxidative stress. These findings are in line with a previous study that CSE overexpression mediated protective effects associated with an increased ERK activation in VSMCs [46].

Bottom Line: UII pretreatment significantly reduced the number of apoptotic cardiomyocytes induced by H2O2; and it partly abolished the increase of pro-apoptotic protein Bax and the decrease of anti-apoptotic protein Bcl-2 in cardiomyocytes induced by H2O2.SiRNA targeted to the urotensin II receptor (UT) greatly inhibited these effects.In conclusion, UII rapidly promoted the phosphorylation of ERK and upregulated CSE level and H2S production, which in turn activated ERK signaling to protect cardiomyocytes from apoptosis under oxidative stress.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China. ghui1975@163.com.

ABSTRACT
Plasma urotensin II (UII) has been observed to be raised in patients with acute myocardial infarction; suggesting a possible cardiac protective role for this peptide. However, the molecular mechanism is unclear. Here, we treated cultured cardiomyocytes with H2O2 to induce oxidative stress; observed the effect of UII on H2O2-induced apoptosis and explored potential mechanisms. UII pretreatment significantly reduced the number of apoptotic cardiomyocytes induced by H2O2; and it partly abolished the increase of pro-apoptotic protein Bax and the decrease of anti-apoptotic protein Bcl-2 in cardiomyocytes induced by H2O2. SiRNA targeted to the urotensin II receptor (UT) greatly inhibited these effects. Further analysis revealed that UII increased the production of hydrogen sulfide (H2S) and the level of cystathionine-γ-lyase (CSE) by activating the ERK signaling in H2O2-treated-cardiomyocytes. Si-CSE or ERK inhibitor not only greatly inhibited the increase in CSE level or the phosphorylation of ERK induced by UII but also reversed anti-apoptosis of UII in H2O2-treated-cadiomyocytes. In conclusion, UII rapidly promoted the phosphorylation of ERK and upregulated CSE level and H2S production, which in turn activated ERK signaling to protect cardiomyocytes from apoptosis under oxidative stress. These results suggest that increased plasma UII level may protect cardiomyocytes at the early-phase of acute myocardial infarction in patients.

No MeSH data available.


Related in: MedlinePlus